2c.bmp (14.1 MB) Hi there,
I want to count the no. of cells in the image and also want to divide them based on their RGB values and count the no. of cells again.
attached is the sample image taken in fluorescence microscopy.

O.k. The image you have shared is an RGB image. In Cellprofiler you can use “colorToGray” module to split your RGB image. Then you can use other primary/secondary object module to segment the nuclei and cells. To segment nuclei & cells you can refer to any of the basic pipeline in this example page.https://cellprofiler.org/examples/